{"gene":"AIFM2","run_date":"2026-06-09T22:02:42","timeline":{"discoveries":[{"year":2002,"finding":"AMID (AIFM2) is a mitochondrion-associated flavoprotein that induces caspase-independent apoptosis independent of p53 and not inhibited by Bcl-2; it lacks a mitochondrial localization sequence but co-localizes with mitochondria as shown by immunofluorescent staining and biochemical fractionation.","method":"Immunofluorescent staining, subcellular fractionation, overexpression in cells, caspase activity assays","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal localization and functional data from overexpression with multiple readouts, single lab","pmids":["11980907"],"is_preprint":false},{"year":2005,"finding":"AMID (AIFM2) is a flavoprotein oxidoreductase that binds the unusual modified cofactor 6-hydroxy-FAD (stoichiometrically), possesses NAD(P)H oxidase activity, and catalyzes NAD(P)H-dependent reduction of cytochrome c and molecular oxygen; NADPH binds ~10-fold more tightly than NADH. AMID also binds DNA without apparent sequence specificity, and coenzyme and DNA binding are mutually exclusive, with DNA binding inducing a major conformational change.","method":"Recombinant protein purification, in vitro enzymatic assays, spectrophotometric cofactor analysis, binding kinetics (Kd determination), DNA-binding assays","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstituted biochemistry with purified recombinant protein, multiple orthogonal in vitro assays, mutagenesis-level cofactor reconstitution, single rigorous study","pmids":["15958387"],"is_preprint":false},{"year":2006,"finding":"The yeast AMID homologue Ndi1p (internal NADH dehydrogenase) induces apoptosis-like cell death when overexpressed, an effect suppressed by increased respiration; Ndi1p overexpression increases mitochondrial ROS production, and disruption of NDI1 decreases ROS and extends chronological lifespan.","method":"Yeast genetics, overexpression/disruption, ROS measurement, chronological lifespan assay","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function and overexpression with multiple cellular phenotype readouts, single lab, yeast ortholog","pmids":["16436509"],"is_preprint":false},{"year":2007,"finding":"DNA binding to AMID (AIF-M2) suppresses its redox activity by preventing NAD(P)H coenzyme binding; binding of DNA and coenzyme is mutually exclusive. DNA binding does not perturb the reduction potential of the 6-hydroxy-FAD cofactor but significantly slows reduction kinetics by NAD(P)H. The DNA-binding domain resides in the C-terminal deletion mutant AIF-M2-(Δ319-613), distinct from AIF whose DNA binds the C-terminal D3 domain.","method":"Solution binding assays, stopped-flow kinetics, redox titrations, deletion mutagenesis, quantitative ligand binding analysis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — multiple orthogonal biochemical methods with mutagenesis, in vitro reconstitution, rigorous quantitative analysis in single study","pmids":["17711848"],"is_preprint":false},{"year":2007,"finding":"AMID protein localizes to the cytoplasm, predominantly at the cytoplasmic side of membranes of various organelles with highest concentration at the Golgi; AMID does not translocate to the nucleus during apoptosis (unlike AIF and endonuclease G), and bioinformatic analysis does not detect nuclear localization signals in AMID.","method":"Live-cell fluorescence imaging, single-cell colocalization analysis, time-lapse imaging during staurosporine-induced apoptosis, bioinformatic prediction, nuclear translocation analysis","journal":"Apoptosis : an international journal on programmed cell death","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — direct live-cell imaging with quantitative colocalization, multiple methods, but finding of no nuclear translocation is a negative result","pmids":["17347867"],"is_preprint":false},{"year":2004,"finding":"The human AMID gene promoter is a direct transcriptional target of p53: p53 binds the AMID promoter in chromatin immunoprecipitation assays, and deletion mutagenesis identified two p53-responsive elements in the promoter.","method":"Luciferase reporter assays, chromatin immunoprecipitation (ChIP), deletion mutagenesis, expression arrays","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus reporter assay with mutagenesis, single lab","pmids":["15273740"],"is_preprint":false},{"year":2006,"finding":"AMID-deficient mice generated by gene targeting are viable and fertile with no spontaneous tumors and similar fibrosarcoma incidence after MCA inoculation compared to wild-type; AMID-/- embryonic fibroblasts show slightly increased resistance to genotoxin-induced growth arrest. These data indicate AMID is not required for normal development or p53-mediated tumor suppression.","method":"Gene targeting (knockout mouse), tumor incidence assays, MEF proliferation and genotoxin-resistance assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean genetic KO in vivo with multiple phenotypic readouts; negative finding for tumor suppression robustly established","pmids":["16186796"],"is_preprint":false},{"year":2008,"finding":"AMID protein is found associated with the plasma membrane fraction of apoptotic cells (identified by MALDI-TOF mass spectrometry from Pisum sativum lectin-affinity chromatography), suggesting possible glycosylation and translocation to the plasma membrane during apoptosis; in Jurkat T-cells, AMID expression increases after apoptosis induction.","method":"Lectin-affinity chromatography, MALDI-TOF mass spectrometry, immunocytochemistry, Western blotting","journal":"Apoptosis : an international journal on programmed cell death","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single approach for membrane localization, no direct functional validation of glycosylation","pmids":["18368494"],"is_preprint":false},{"year":2015,"finding":"AMID (AIFM2) functions as a rotenone-sensitive NADH:ubiquinone oxidoreductase (NDH-2): when reconstituted into bacterial or mitochondrial membranes, N-terminally tagged AMID displays substantial NADH:O2 activity, supports NADH-linked proton pumping, and rescues growth of E. coli lacking both complex I and NDH-2. C-terminally tagged AMID and NADH-binding site mutants fail to show these activities.","method":"Membrane reconstitution, NADH:O2 activity assays, proton pumping assays, E. coli complementation (growth rescue), site-directed mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro reconstitution, mutagenesis, and heterologous complementation with multiple orthogonal assays in one study","pmids":["26063804"],"is_preprint":false},{"year":2012,"finding":"Intracellularly transported adenosine induces caspase-independent apoptosis in MCF-7 breast cancer cells by promoting translocation of AMID from the cytosol to the nucleus; AMID knockdown abrogates adenosine-induced cell death.","method":"siRNA knockdown, immunofluorescence (AMID localization), caspase activity assays, cell viability assays, adenosine transporter inhibitor studies","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — knockdown with rescue-relevant pathway placement and localization data, single lab","pmids":["22388174"],"is_preprint":false},{"year":2012,"finding":"A3 adenosine receptor activation mediates apoptosis in RCC4-VHL human renal cancer cells by upregulating AMID mRNA and protein expression and promoting AMID translocation from cytosol to nucleus; knockdown of either A3 receptor or AMID inhibits adenosine-induced apoptosis.","method":"siRNA knockdown, RT-PCR, Western blotting, immunofluorescence, MTT assay, TUNEL staining","journal":"The Journal of urology","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal knockdown epistasis establishes pathway position, single lab","pmids":["23174235"],"is_preprint":false},{"year":2020,"finding":"AIFM2/FSP1 blocks ferroptosis through ESCRT-III recruitment to the plasma membrane, activating a membrane repair mechanism involving membrane budding and fission; this mechanism is independent of ubiquinol/coenzyme Q10. Genetic inhibition of the AIFM2-ESCRT-III pathway increases anticancer activity of sorafenib in a xenograft mouse model.","method":"Genetic inhibition, xenograft mouse model, ferroptosis induction assays (erastin, sorafenib, RSL3), ESCRT-III pathway analysis","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — in vitro and in vivo genetic experiments with mechanistic pathway placement, single lab","pmids":["31964528"],"is_preprint":false},{"year":2020,"finding":"Aifm2 is a lipid droplet-associated NADH oxidoreductase enriched in brown adipose tissue (BAT) that, upon cold or β-adrenergic stimulation, associates with the outer side of the mitochondrial inner membrane. Acting as a mammalian external NADH dehydrogenase (NDE)-like enzyme, Aifm2 oxidizes NADH to maintain high cytosolic NAD+ levels supporting robust glycolysis and transfers electrons to the electron transport chain (ETC) to fuel thermogenesis. Knockout of Aifm2 impairs cold- and diet-induced thermogenesis, oxygen consumption, and uncoupled respiration.","method":"Subcellular fractionation, lipid droplet isolation, cold/β-adrenergic stimulation experiments, NAD+/NADH measurements, oxygen consumption assays, KO mouse model, diet-induced obesity model","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — multiple orthogonal biochemical and genetic methods in vivo and in vitro, KO mouse with defined metabolic phenotypes, subcellular localization with functional consequence","pmids":["31952989"],"is_preprint":false},{"year":2023,"finding":"FSP1 (AIFM2) inhibition by iFSP1 is species-specific; using cellular, biochemical, and computational methods, a species-specific aromatic architecture was identified as critical for target engagement with the human isoform.","method":"Biochemical inhibition assays, computational modeling, cellular ferroptosis assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (biochemical + computational + cellular), single lab","pmids":["37080964"],"is_preprint":false},{"year":2020,"finding":"ATF6 promotes AIFM2 transcription by binding to both the p53 and AIFM2 promoters; the ATF6/p53/AIFM2 pathway drives acinar cell apoptosis in severe acute pancreatitis. ATF6 knockout suppresses AIFM2 expression and reduces apoptosis, while AIFM2 overexpression reestablishes pathological disorder in ATF6-knockout SAP tissues.","method":"ChIP-qPCR, luciferase reporter assay, ATF6 knockout mouse model, adenovirus-mediated shRNA knockdown/overexpression, Western blotting, immunofluorescence","journal":"Theranostics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and reporter assays establish direct promoter binding, genetic epistasis in vivo, single lab","pmids":["32724472"],"is_preprint":false},{"year":2025,"finding":"USP7 deubiquitinates and stabilizes the transcription factor JunD, which in turn promotes transcription of AIFM2 (FSP1), enhancing ferroptosis resistance in melanoma. USP7 inhibition leads to JunD degradation and reduced AIFM2 levels, sensitizing melanoma cells to ferroptosis in vitro and in murine xenograft models.","method":"Deubiquitinase identification, Co-IP, Western blotting, gene knockdown/inhibitor experiments, xenograft mouse model, ferroptosis assays","journal":"The Journal of investigative dermatology","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — epistasis chain established with in vitro and in vivo confirmation, single lab","pmids":["40187679"],"is_preprint":false},{"year":2023,"finding":"AIFM2 promotes HCC metastasis by activating SIRT1/PGC-1α signaling, increasing mitochondrial biogenesis and oxidative phosphorylation; AIFM2 knockdown impairs HCC metastasis in vitro and in vivo, while overexpression enhances it.","method":"shRNA knockdown, overexpression, in vitro migration/invasion assays, in vivo metastasis models, Western blotting for SIRT1/PGC-1α pathway components","journal":"Oncogenesis","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — gain- and loss-of-function with in vivo validation and pathway identification, single lab","pmids":["37735151"],"is_preprint":false},{"year":2025,"finding":"IGF2BP1 stabilizes AIFM2 mRNA via m6A modification, promoting AIFM2 expression; AIFM2 knockdown suppresses glycolysis and enhances ferroptosis in HCC cells, while overexpression has the opposite effect. IGF2BP1 knockdown effects on glycolysis and ferroptosis are rescued by AIFM2 overexpression.","method":"MeRIP assay (m6A detection), mRNA stability assay (Actinomycin D), RIP, siRNA knockdown, overexpression, ferroptosis marker measurement (ROS, iron, MDA/GSH/SOD), glycolysis assays, xenograft/metastasis models","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct RNA modification assay (MeRIP) combined with functional rescue experiments, single lab","pmids":["39971223"],"is_preprint":false},{"year":2015,"finding":"HUHS1015 (naftopidil analogue) induces caspase-independent apoptosis in MKN28 gastric cancer cells associated with increased nuclear localization of AMID, without changing AMID mRNA or total protein levels, indicating post-translational redistribution.","method":"Immunofluorescence (nuclear AMID localization), caspase activity assays, cell cycle analysis, flow cytometry, RT-PCR, Western blotting","journal":"Anti-cancer agents in medicinal chemistry","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, correlative localization finding without mechanistic dissection of the translocation mechanism","pmids":["25244912"],"is_preprint":false},{"year":2011,"finding":"Adenosine upregulates AMID expression and promotes translocation of AMID from cytosol to nucleus in HuH-7 hepatoma cells; AMID knockdown inhibits adenosine-induced caspase-independent apoptosis, and AMID overexpression enhances it.","method":"RT-PCR, immunofluorescence, time-lapse GFP imaging, siRNA knockdown, overexpression, TUNEL staining, flow cytometry","journal":"Cellular physiology and biochemistry","confidence":"Medium","confidence_rationale":"Tier 2–3 / Moderate — reciprocal gain and loss of function with localization data, single lab","pmids":["21325820"],"is_preprint":false}],"current_model":"AIFM2 (AMID/FSP1) is a flavoprotein NADH oxidoreductase that binds 6-hydroxy-FAD and functions as an external NADH dehydrogenase (NDH-2): it oxidizes cytosolic NADH to maintain NAD+ levels supporting glycolysis and feeds electrons to the mitochondrial electron transport chain; its transcription is directly activated by p53 and by ATF6 via p53, and its expression is stabilized post-transcriptionally via IGF2BP1-mediated m6A modification; in ferroptosis, AIFM2/FSP1 suppresses lipid peroxidation-driven cell death through plasma membrane repair via ESCRT-III recruitment and through its reductase activity; DNA binding suppresses its oxidoreductase activity by excluding NAD(P)H; in apoptotic contexts, AMID undergoes cytosol-to-nucleus translocation to drive caspase-independent cell death; and USP7-stabilized JunD directly promotes AIFM2 transcription to confer ferroptosis resistance."},"narrative":{"mechanistic_narrative":"AIFM2 (AMID/FSP1) is a flavoprotein NAD(P)H oxidoreductase that couples cytosolic redox metabolism to cell-fate decisions [PMID:15958387, PMID:31952989]. As a purified enzyme it binds the unusual cofactor 6-hydroxy-FAD stoichiometrically, possesses NAD(P)H oxidase activity, and catalyzes NAD(P)H-dependent reduction of cytochrome c and molecular oxygen, with NADPH binding more tightly than NADH [PMID:15958387]. When reconstituted into membranes it behaves as a rotenone-sensitive NADH:ubiquinone oxidoreductase (NDH-2), supporting NADH:O2 activity and NADH-linked proton pumping and rescuing E. coli lacking complex I and NDH-2 [PMID:26063804]. Consistent with this enzymology, Aifm2 acts in vivo as a mammalian external NADH dehydrogenase that oxidizes cytosolic NADH to maintain NAD+ for glycolysis and feeds electrons to the electron transport chain; in brown adipose tissue it is lipid-droplet associated and relocates to the outer face of the mitochondrial inner membrane upon cold or β-adrenergic stimulation, and its loss impairs thermogenesis and uncoupled respiration [PMID:31952989]. AIFM2 also binds DNA without sequence specificity, and DNA and coenzyme binding are mutually exclusive: DNA binding induces a major conformational change and suppresses redox activity by excluding NAD(P)H [PMID:15958387, PMID:17711848]. In ferroptosis, AIFM2/FSP1 acts as a suppressor of lipid-peroxidation-driven death, including through recruitment of ESCRT-III to the plasma membrane to drive membrane repair independently of coenzyme Q10 [PMID:31964528]. Independently, AMID can drive caspase-independent apoptosis: it is a mitochondrion-associated flavoprotein that triggers death not inhibited by Bcl-2 [PMID:11980907], and in several cancer contexts adenosine signaling upregulates AMID and promotes its translocation from cytosol to nucleus to execute cell death [PMID:22388174, PMID:21325820]. Its expression is controlled transcriptionally — the promoter is a direct p53 target [PMID:15273740], is activated by ATF6 acting via p53 in acute pancreatitis [PMID:32724472], and is driven by USP7-stabilized JunD to confer ferroptosis resistance in melanoma [PMID:40187679] — and post-transcriptionally through IGF2BP1-mediated m6A stabilization of its mRNA, which supports glycolysis and ferroptosis resistance [PMID:39971223]. Genetic deletion of Aifm2 in mice is compatible with normal development and is dispensable for p53-mediated tumor suppression [PMID:16186796].","teleology":[{"year":2002,"claim":"Established AMID as a death-inducing mitochondrion-associated flavoprotein, raising the question of how a protein lacking a mitochondrial targeting sequence executes caspase-independent apoptosis.","evidence":"Overexpression, immunofluorescence, subcellular fractionation and caspase assays in cells","pmids":["11980907"],"confidence":"Medium","gaps":["Death effector mechanism not defined","Overexpression-based; endogenous role unaddressed"]},{"year":2004,"claim":"Placed AMID under direct transcriptional control of p53, linking its expression to tumor-suppressor signaling.","evidence":"ChIP, luciferase reporter and deletion mutagenesis of the human AMID promoter","pmids":["15273740"],"confidence":"Medium","gaps":["Whether p53-driven AMID contributes to apoptosis in vivo untested","Single cell-context"]},{"year":2005,"claim":"Defined AMID as a bona fide oxidoreductase, identifying its 6-hydroxy-FAD cofactor and NAD(P)H oxidase activity, and revealing mutually exclusive DNA and coenzyme binding.","evidence":"Recombinant protein, in vitro enzymatic and binding-kinetics assays, DNA-binding assays","pmids":["15958387"],"confidence":"High","gaps":["Physiological electron acceptor in cells not established","Functional meaning of DNA binding unresolved"]},{"year":2006,"claim":"Genetic knockout showed AMID is dispensable for development and p53-mediated tumor suppression, decoupling its in vitro death activity from an essential physiological requirement.","evidence":"Knockout mouse with tumor-incidence and MEF genotoxin-resistance assays","pmids":["16186796"],"confidence":"High","gaps":["Does not exclude context-specific or redundant functions","Metabolic phenotypes not examined"]},{"year":2007,"claim":"Resolved the mechanism by which DNA binding regulates AMID enzymology — DNA slows NAD(P)H reduction and a C-terminal domain mediates DNA binding — establishing a redox/DNA switch.","evidence":"Stopped-flow kinetics, redox titrations and deletion mutagenesis","pmids":["17711848"],"confidence":"High","gaps":["In vivo relevance of DNA-mediated redox suppression unknown","DNA-binding domain structure not solved"]},{"year":2007,"claim":"Localization imaging placed AMID predominantly at cytoplasmic membrane faces (highest at Golgi) and reported no apoptotic nuclear translocation, contesting an AIF-like nuclear death mechanism.","evidence":"Live-cell imaging and time-lapse colocalization during staurosporine-induced apoptosis","pmids":["17347867"],"confidence":"Medium","gaps":["Negative nuclear-translocation result conflicts with later cancer studies","Stimulus- and cell-type-specific behavior not reconciled"]},{"year":2006,"claim":"The yeast homologue Ndi1p linked NADH dehydrogenase activity to ROS-dependent death and lifespan, framing the redox-versus-death duality conserved across the family.","evidence":"Yeast overexpression/disruption, ROS measurement, chronological lifespan assays","pmids":["16436509"],"confidence":"Medium","gaps":["Ortholog inference, not direct AIFM2 data","Mechanism connecting ROS to death not dissected"]},{"year":2011,"claim":"Adenosine signaling was shown to upregulate AMID and drive its cytosol-to-nucleus translocation in caspase-independent apoptosis, defining an upstream death-activation pathway.","evidence":"RT-PCR, GFP time-lapse imaging, siRNA and overexpression with TUNEL in hepatoma cells","pmids":["21325820"],"confidence":"Medium","gaps":["Nuclear targeting mechanism unknown (no NLS reported)","Nuclear death effector activity undefined"]},{"year":2012,"claim":"Extended the adenosine-AMID translocation pathway across breast and renal cancer cells and identified A3 adenosine receptor as an upstream node, generalizing the death mechanism.","evidence":"siRNA knockdown, immunofluorescence, viability/TUNEL assays and transporter-inhibitor studies","pmids":["22388174","23174235"],"confidence":"Medium","gaps":["Direct molecular trigger of translocation unresolved","Reconciliation with negative nuclear-translocation report needed"]},{"year":2015,"claim":"Reconstitution defined AMID as an NDH-2-type NADH:ubiquinone oxidoreductase capable of proton pumping and heterologous complementation, anchoring its bioenergetic function.","evidence":"Membrane reconstitution, NADH:O2 and proton-pumping assays, E. coli complementation, site-directed mutagenesis","pmids":["26063804"],"confidence":"High","gaps":["Topology and physiological membrane association in mammalian cells not fully resolved here","Connection to apoptotic function unclear"]},{"year":2020,"claim":"Identified Aifm2 as a lipid-droplet/mitochondrial external NADH dehydrogenase that sustains cytosolic NAD+ for glycolysis and fuels thermogenesis, defining its core metabolic physiology in vivo.","evidence":"Subcellular fractionation, NAD+/NADH and oxygen-consumption assays, KO mouse and diet-induced obesity models","pmids":["31952989"],"confidence":"High","gaps":["Regulation of stimulus-dependent relocalization mechanistically incomplete","Relationship to ferroptosis pool of AIFM2 not addressed"]},{"year":2020,"claim":"Revealed a CoQ-independent ferroptosis-suppressing mechanism in which AIFM2/FSP1 recruits ESCRT-III for plasma-membrane repair, broadening its anti-ferroptotic role beyond reductase activity.","evidence":"Genetic inhibition, ferroptosis induction assays and xenograft model","pmids":["31964528"],"confidence":"Medium","gaps":["Molecular link between AIFM2 and ESCRT-III recruitment undefined","Relative contribution of repair versus reductase activity unquantified"]},{"year":2020,"claim":"Showed ATF6 transactivates AIFM2 via p53 to drive acinar apoptosis, embedding AIFM2 in an ER-stress transcriptional axis in pancreatitis.","evidence":"ChIP-qPCR, reporter assays, ATF6 KO mouse and shRNA/overexpression epistasis","pmids":["32724472"],"confidence":"Medium","gaps":["Direct versus p53-dependent contribution of ATF6 not fully separated","Single disease context"]},{"year":2023,"claim":"Characterized species-specific pharmacology of the FSP1 inhibitor iFSP1, identifying an aromatic architecture required for human target engagement, informing therapeutic targeting.","evidence":"Biochemical inhibition, computational modeling and cellular ferroptosis assays","pmids":["37080964"],"confidence":"Medium","gaps":["Structural binding mode not crystallographically resolved","In vivo efficacy across species not tested"]},{"year":2023,"claim":"Linked AIFM2 to a pro-metastatic program via SIRT1/PGC-1α-driven mitochondrial biogenesis and oxidative phosphorylation, connecting its redox role to cancer aggressiveness.","evidence":"shRNA/overexpression, migration/invasion and in vivo metastasis assays in HCC","pmids":["37735151"],"confidence":"Medium","gaps":["Mechanism by which AIFM2 activates SIRT1/PGC-1α unclear","Whether reductase activity is required not tested"]},{"year":2025,"claim":"Defined transcriptional and post-transcriptional control of AIFM2 in ferroptosis resistance — USP7–JunD transactivation and IGF2BP1 m6A mRNA stabilization — coupling AIFM2 expression to glycolysis and cancer cell survival.","evidence":"Co-IP, DUB identification, MeRIP/RIP, mRNA-stability and rescue assays with xenograft models","pmids":["40187679","39971223"],"confidence":"Medium","gaps":["Whether these regulators act on the same AIFM2 pool as the metabolic/ferroptosis function unclear","Direct binding of JunD to AIFM2 promoter mapped only in melanoma"]},{"year":null,"claim":"It remains unresolved how a single flavoprotein partitions between its bioenergetic NADH-dehydrogenase role, its membrane-repair/ferroptosis-suppressing role, and its pro-apoptotic nuclear-translocation behavior, and what governs that partitioning.","evidence":"No timeline discovery reconciles the metabolic, ferroptotic, and apoptotic activities mechanistically","pmids":[],"confidence":"Low","gaps":["No structural model integrating DNA-binding, membrane-association and reductase states","Conflicting nuclear-translocation reports unreconciled","Trigger for context-specific functional switching unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016491","term_label":"oxidoreductase activity","supporting_discovery_ids":[1,8,12]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[1,3]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[3]}],"localization":[{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[0,12]},{"term_id":"GO:0005811","term_label":"lipid droplet","supporting_discovery_ids":[12]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[11]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[4,9]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[4]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[9,19]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,11]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[12,8]}],"complexes":[],"partners":["ESCRT-III"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BRQ8","full_name":"Ferroptosis suppressor protein 1","aliases":["Apoptosis-inducing factor homologous mitochondrion-associated inducer of death","AMID","p53-responsive gene 3 protein"],"length_aa":373,"mass_kda":40.5,"function":"A NAD(P)H-dependent oxidoreductase that acts as a key inhibitor of ferroptosis (PubMed:31634899, PubMed:31634900, PubMed:35922516, PubMed:36788244, PubMed:37957306, PubMed:39881208, PubMed:37739993, PubMed:39160155, PubMed:38414669, PubMed:40233740). At the plasma membrane, catalyzes reduction of coenzyme Q/ubiquinone-10 to ubiquinol-10, a lipophilic radical-trapping antioxidant that prevents lipid oxidative damage and consequently ferroptosis (PubMed:31634899, PubMed:31634900, PubMed:37739993, PubMed:39160155, PubMed:38414669, PubMed:40233740). Acts in parallel to GPX4 to suppress phospholipid peroxidation and ferroptosis (PubMed:31634899, PubMed:31634900). This anti-ferroptotic function is independent of cellular glutathione levels (PubMed:31634899, PubMed:31634900). Also acts as a potent radical-trapping antioxidant by mediating warfarin-resistant vitamin K reduction in the canonical vitamin K cycle: catalyzes NAD(P)H-dependent reduction of vitamin K (phylloquinone, menaquinone-4 and menadione) to hydroquinone forms (PubMed:35922516, PubMed:36788244). Hydroquinones act as potent radical-trapping antioxidants inhibitor of phospholipid peroxidation and ferroptosis (PubMed:35922516). May play a role in mitochondrial stress signaling (PubMed:26689472). Upon oxidative stress, associates with the lipid peroxidation end product 4-hydroxy-2-nonenal (HNE) forming a lipid adduct devoid of oxidoreductase activity, which then translocates from mitochondria into the nucleus triggering DNA damage and cell death (PubMed:26689472). Capable of DNA binding in a non-sequence specific way (PubMed:15958387)","subcellular_location":"Lipid droplet; Cell membrane; Cytoplasm; Mitochondrion membrane; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9BRQ8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/AIFM2","classification":"Not Classified","n_dependent_lines":18,"n_total_lines":1208,"dependency_fraction":0.014900662251655629},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/AIFM2","total_profiled":1310},"omim":[{"mim_id":"621004","title":"AUTOIMMUNE DISEASE, MULTISYSTEM, INFANTILE-ONSET, 4; ADMIO4","url":"https://www.omim.org/entry/621004"},{"mim_id":"605159","title":"APOPTOSIS-INDUCING FACTOR, MITOCHONDRIA-ASSOCIATED, 2; AIFM2","url":"https://www.omim.org/entry/605159"},{"mim_id":"600244","title":"PROGRAMMED CELL DEATH 1; PDCD1","url":"https://www.omim.org/entry/600244"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Vesicles","reliability":"Supported"},{"location":"Plasma membrane","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/AIFM2"},"hgnc":{"alias_symbol":["FLJ14497","PRG3","FSP1"],"prev_symbol":["AMID"]},"alphafold":{"accession":"Q9BRQ8","domains":[{"cath_id":"3.50.50.60","chopping":"15-43_81-114_246-325","consensus_level":"high","plddt":97.2236,"start":15,"end":325},{"cath_id":"3.50.50.60","chopping":"126-245_327-362","consensus_level":"medium","plddt":97.2213,"start":126,"end":362}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRQ8","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRQ8-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BRQ8-F1-predicted_aligned_error_v6.png","plddt_mean":95.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=AIFM2","jax_strain_url":"https://www.jax.org/strain/search?query=AIFM2"},"sequence":{"accession":"Q9BRQ8","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BRQ8.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BRQ8/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BRQ8"}},"corpus_meta":[{"pmid":"16224497","id":"PMC_16224497","title":"Axon degeneration mechanisms: commonality amid diversity.","date":"2005","source":"Nature reviews. Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/16224497","citation_count":677,"is_preprint":false},{"pmid":"18852696","id":"PMC_18852696","title":"Biological principles of microRNA-mediated regulation: shared themes amid diversity.","date":"2008","source":"Nature reviews. Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/18852696","citation_count":646,"is_preprint":false},{"pmid":"11980907","id":"PMC_11980907","title":"AMID, an apoptosis-inducing factor-homologous mitochondrion-associated protein, induces caspase-independent apoptosis.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/11980907","citation_count":216,"is_preprint":false},{"pmid":"35093310","id":"PMC_35093310","title":"Emergence of SARS-CoV-2 Omicron (B.1.1.529) variant, salient features, high global health concerns and strategies to counter it amid ongoing COVID-19 pandemic.","date":"2022","source":"Environmental research","url":"https://pubmed.ncbi.nlm.nih.gov/35093310","citation_count":194,"is_preprint":false},{"pmid":"31964528","id":"PMC_31964528","title":"AIFM2 blocks ferroptosis independent of ubiquinol metabolism.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31964528","citation_count":188,"is_preprint":false},{"pmid":"33719302","id":"PMC_33719302","title":"Global Regulation of Genetically Modified Crops Amid the Gene Edited Crop Boom - A Review.","date":"2021","source":"Frontiers in plant science","url":"https://pubmed.ncbi.nlm.nih.gov/33719302","citation_count":186,"is_preprint":false},{"pmid":"18053752","id":"PMC_18053752","title":"The role of protein synthesis in memory consolidation: progress amid decades of debate.","date":"2007","source":"Neurobiology of learning and memory","url":"https://pubmed.ncbi.nlm.nih.gov/18053752","citation_count":181,"is_preprint":false},{"pmid":"11004460","id":"PMC_11004460","title":"The mitochondrial cyanide-resistant oxidase: structural conservation amid regulatory diversity.","date":"2000","source":"Biochimica et biophysica acta","url":"https://pubmed.ncbi.nlm.nih.gov/11004460","citation_count":151,"is_preprint":false},{"pmid":"29118215","id":"PMC_29118215","title":"Adolescence and Reward: Making Sense of Neural and Behavioral Changes Amid the Chaos.","date":"2017","source":"The Journal of neuroscience : the official journal of the Society for Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/29118215","citation_count":138,"is_preprint":false},{"pmid":"28356329","id":"PMC_28356329","title":"Effectors of Filamentous Plant Pathogens: Commonalities amid Diversity.","date":"2017","source":"Microbiology and molecular biology reviews : MMBR","url":"https://pubmed.ncbi.nlm.nih.gov/28356329","citation_count":137,"is_preprint":false},{"pmid":"26496035","id":"PMC_26496035","title":"Cancer stem cell targeted therapy: progress amid controversies.","date":"2015","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/26496035","citation_count":122,"is_preprint":false},{"pmid":"27150399","id":"PMC_27150399","title":"The landscape of DNA methylation amid a perfect storm of autism aetiologies.","date":"2016","source":"Nature reviews. Neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/27150399","citation_count":120,"is_preprint":false},{"pmid":"16436509","id":"PMC_16436509","title":"Yeast AMID homologue Ndi1p displays respiration-restricted apoptotic activity and is involved in chronological aging.","date":"2006","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/16436509","citation_count":118,"is_preprint":false},{"pmid":"31952989","id":"PMC_31952989","title":"Aifm2, a NADH Oxidase, Supports Robust Glycolysis and Is Required for Cold- and Diet-Induced Thermogenesis.","date":"2020","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/31952989","citation_count":116,"is_preprint":false},{"pmid":"28827147","id":"PMC_28827147","title":"RING-Between-RING E3 Ligases: Emerging Themes amid the Variations.","date":"2017","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/28827147","citation_count":116,"is_preprint":false},{"pmid":"29378836","id":"PMC_29378836","title":"Plant cell surface receptor-mediated signaling - a common theme amid diversity.","date":"2018","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/29378836","citation_count":111,"is_preprint":false},{"pmid":"15958387","id":"PMC_15958387","title":"The human apoptosis-inducing protein AMID is an oxidoreductase with a modified flavin cofactor and DNA binding activity.","date":"2005","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/15958387","citation_count":102,"is_preprint":false},{"pmid":"26063804","id":"PMC_26063804","title":"Apoptosis-inducing Factor (AIF) and Its Family Member Protein, AMID, Are Rotenone-sensitive NADH:Ubiquinone Oxidoreductases (NDH-2).","date":"2015","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/26063804","citation_count":94,"is_preprint":false},{"pmid":"32299148","id":"PMC_32299148","title":"Considerations for Obesity, Vitamin D, and Physical Activity Amid the COVID-19 Pandemic.","date":"2020","source":"Obesity (Silver Spring, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/32299148","citation_count":91,"is_preprint":false},{"pmid":"36030585","id":"PMC_36030585","title":"Omicron variant (B.1.1.529) and its sublineages: What do we know so far amid the emergence of recombinant variants of SARS-CoV-2?","date":"2022","source":"Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie","url":"https://pubmed.ncbi.nlm.nih.gov/36030585","citation_count":83,"is_preprint":false},{"pmid":"7530487","id":"PMC_7530487","title":"Cytogenetic analysis of 63 non-small cell lung carcinomas: recurrent chromosome alterations amid frequent and widespread genomic upheaval.","date":"1994","source":"Genes, chromosomes & cancer","url":"https://pubmed.ncbi.nlm.nih.gov/7530487","citation_count":83,"is_preprint":false},{"pmid":"32724472","id":"PMC_32724472","title":"ATF6 aggravates acinar cell apoptosis and injury by regulating p53/AIFM2 transcription in Severe Acute Pancreatitis.","date":"2020","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/32724472","citation_count":78,"is_preprint":false},{"pmid":"20089121","id":"PMC_20089121","title":"Self-recruitment and sweepstakes reproduction amid extensive gene flow in a coral-reef fish.","date":"2010","source":"Molecular ecology","url":"https://pubmed.ncbi.nlm.nih.gov/20089121","citation_count":69,"is_preprint":false},{"pmid":"26733718","id":"PMC_26733718","title":"TSPO: kaleidoscopic 18-kDa amid biochemical pharmacology, control and targeting of mitochondria.","date":"2016","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/26733718","citation_count":65,"is_preprint":false},{"pmid":"34668628","id":"PMC_34668628","title":"CircGFRA1 facilitates the malignant progression of HER-2-positive breast cancer via acting as a sponge of miR-1228 and enhancing AIFM2 expression.","date":"2021","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34668628","citation_count":61,"is_preprint":false},{"pmid":"38424189","id":"PMC_38424189","title":"Intestinal stem cells: guardians of homeostasis in health and aging amid environmental challenges.","date":"2024","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38424189","citation_count":59,"is_preprint":false},{"pmid":"37080964","id":"PMC_37080964","title":"Molecular characterization of AIFM2/FSP1 inhibition by iFSP1-like molecules.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/37080964","citation_count":47,"is_preprint":false},{"pmid":"15273740","id":"PMC_15273740","title":"AMID is a p53-inducible gene downregulated in tumors.","date":"2004","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/15273740","citation_count":46,"is_preprint":false},{"pmid":"17711848","id":"PMC_17711848","title":"DNA binding suppresses human AIF-M2 activity and provides a connection between redox chemistry, reactive oxygen species, and apoptosis.","date":"2007","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/17711848","citation_count":45,"is_preprint":false},{"pmid":"27392435","id":"PMC_27392435","title":"Activation of AIFM2 enhances apoptosis of human lung cancer cells undergoing toxicological stress.","date":"2016","source":"Toxicology letters","url":"https://pubmed.ncbi.nlm.nih.gov/27392435","citation_count":43,"is_preprint":false},{"pmid":"32336945","id":"PMC_32336945","title":"Distinct spread of DNA and RNA viruses among mammals amid prominent role of domestic species.","date":"2019","source":"Global ecology and biogeography : a journal of macroecology","url":"https://pubmed.ncbi.nlm.nih.gov/32336945","citation_count":43,"is_preprint":false},{"pmid":"20036252","id":"PMC_20036252","title":"Specific structural features of the N-acetylmuramoyl-L-alanine amidase AmiD from Escherichia coli and mechanistic implications for enzymes of this family.","date":"2009","source":"Journal of molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/20036252","citation_count":41,"is_preprint":false},{"pmid":"19277981","id":"PMC_19277981","title":"Differences amid bone marrow and cord blood hematopoietic stem/progenitor cell division kinetics.","date":"2009","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/19277981","citation_count":36,"is_preprint":false},{"pmid":"17347867","id":"PMC_17347867","title":"Bioinformatic and image analyses of the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID during apoptosis in human cells.","date":"2007","source":"Apoptosis : an international journal on programmed cell death","url":"https://pubmed.ncbi.nlm.nih.gov/17347867","citation_count":36,"is_preprint":false},{"pmid":"34324556","id":"PMC_34324556","title":"Older adults with non-communicable chronic conditions and their health care access amid COVID-19 pandemic in Bangladesh: Findings from a cross-sectional study.","date":"2021","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/34324556","citation_count":36,"is_preprint":false},{"pmid":"37771779","id":"PMC_37771779","title":"5-aminolevulinate and CHIL3/CHI3L1 treatment amid ischemia aids liver metabolism and reduces ischemia-reperfusion injury.","date":"2023","source":"Theranostics","url":"https://pubmed.ncbi.nlm.nih.gov/37771779","citation_count":35,"is_preprint":false},{"pmid":"28534937","id":"PMC_28534937","title":"The inhibitory effect of MEG3/miR-214/AIFM2 axis on the growth of T-cell lymphoblastic lymphoma.","date":"2017","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/28534937","citation_count":35,"is_preprint":false},{"pmid":"16186796","id":"PMC_16186796","title":"The p53-inducible apoptotic protein AMID is not required for normal development and tumor suppression.","date":"2006","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/16186796","citation_count":35,"is_preprint":false},{"pmid":"22388174","id":"PMC_22388174","title":"Intracellularly transported adenosine induces apoptosis in [corrected] MCF-7 human breast cancer cells by accumulating AMID in the nucleus.","date":"2012","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/22388174","citation_count":32,"is_preprint":false},{"pmid":"20943625","id":"PMC_20943625","title":"Soft tissue sarcoma: from molecular diagnosis to selection of treatment. Pathological diagnosis of soft tissue sarcoma amid molecular biology and targeted therapies.","date":"2010","source":"Annals of oncology : official journal of the European Society for Medical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/20943625","citation_count":31,"is_preprint":false},{"pmid":"40437092","id":"PMC_40437092","title":"Domesticated cannabinoid synthases amid a wild mosaic cannabis pangenome.","date":"2025","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/40437092","citation_count":30,"is_preprint":false},{"pmid":"36696432","id":"PMC_36696432","title":"Reinforcement amid genetic diversity in the Candida albicans biofilm regulatory network.","date":"2023","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/36696432","citation_count":30,"is_preprint":false},{"pmid":"26230828","id":"PMC_26230828","title":"Physical Limit to Concentration Sensing Amid Spurious Ligands.","date":"2015","source":"Physical review letters","url":"https://pubmed.ncbi.nlm.nih.gov/26230828","citation_count":30,"is_preprint":false},{"pmid":"18368494","id":"PMC_18368494","title":"AMID: new insights on its intracellular localization and expression at apoptosis.","date":"2008","source":"Apoptosis : an international journal on programmed cell death","url":"https://pubmed.ncbi.nlm.nih.gov/18368494","citation_count":27,"is_preprint":false},{"pmid":"27460344","id":"PMC_27460344","title":"Commonality amid diversity: Multi-study proteomic identification of conserved disease mechanisms in spinal muscular atrophy.","date":"2016","source":"Neuromuscular disorders : NMD","url":"https://pubmed.ncbi.nlm.nih.gov/27460344","citation_count":27,"is_preprint":false},{"pmid":"16427003","id":"PMC_16427003","title":"Modified view of tRNA: stability amid sequence diversity.","date":"2006","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/16427003","citation_count":24,"is_preprint":false},{"pmid":"23174235","id":"PMC_23174235","title":"A3 adenosine receptor mediates apoptosis in in vitro RCC4-VHL human renal cancer cells by up-regulating AMID expression.","date":"2012","source":"The Journal of urology","url":"https://pubmed.ncbi.nlm.nih.gov/23174235","citation_count":23,"is_preprint":false},{"pmid":"19237650","id":"PMC_19237650","title":"Substrate-induced inactivation of the Escherichia coli AmiD N-acetylmuramoyl-L-alanine amidase highlights a new strategy to inhibit this class of enzyme.","date":"2009","source":"Antimicrobial agents and chemotherapy","url":"https://pubmed.ncbi.nlm.nih.gov/19237650","citation_count":23,"is_preprint":false},{"pmid":"10022978","id":"PMC_10022978","title":"Analysis of two cosmid clones from chromosome 4 of Drosophila melanogaster reveals two new genes amid an unusual arrangement of repeated sequences.","date":"1999","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/10022978","citation_count":22,"is_preprint":false},{"pmid":"16950880","id":"PMC_16950880","title":"Accurately quantifying low-abundant targets amid similar sequences by revealing hidden correlations in oligonucleotide microarray data.","date":"2006","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/16950880","citation_count":22,"is_preprint":false},{"pmid":"24478800","id":"PMC_24478800","title":"Adaptive genetic markers discriminate migratory runs of Chinook salmon (Oncorhynchus tshawytscha) amid continued gene flow.","date":"2013","source":"Evolutionary applications","url":"https://pubmed.ncbi.nlm.nih.gov/24478800","citation_count":22,"is_preprint":false},{"pmid":"30824938","id":"PMC_30824938","title":"Brassicaceae flowers: diversity amid uniformity.","date":"2019","source":"Journal of experimental botany","url":"https://pubmed.ncbi.nlm.nih.gov/30824938","citation_count":20,"is_preprint":false},{"pmid":"21325820","id":"PMC_21325820","title":"AMID mediates adenosine-induced caspase-independent HuH-7 cell apoptosis.","date":"2011","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/21325820","citation_count":20,"is_preprint":false},{"pmid":"34320263","id":"PMC_34320263","title":"Facing the wrath of enigmatic mutations: a review on the emergence of severe acute respiratory syndrome coronavirus 2 variants amid coronavirus disease-19 pandemic.","date":"2021","source":"Environmental microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/34320263","citation_count":20,"is_preprint":false},{"pmid":"22854535","id":"PMC_22854535","title":"Adenosine induces apoptosis in SBC-3 human lung cancer cells through A(3) adenosine receptor-dependent AMID upregulation.","date":"2012","source":"Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/22854535","citation_count":20,"is_preprint":false},{"pmid":"35974010","id":"PMC_35974010","title":"Impedance-based forecasting of lithium-ion battery performance amid uneven usage.","date":"2022","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/35974010","citation_count":20,"is_preprint":false},{"pmid":"33587724","id":"PMC_33587724","title":"Increased Intracranial Hemorrhage Amid Elevated Inflammatory Markers in Those With COVID-19 Supported With Extracorporeal Membrane Oxygenation.","date":"2021","source":"Shock (Augusta, Ga.)","url":"https://pubmed.ncbi.nlm.nih.gov/33587724","citation_count":20,"is_preprint":false},{"pmid":"38244593","id":"PMC_38244593","title":"Circ0060467 sponges miR-6805 to promote hepatocellular carcinoma progression through regulating AIFM2 and GPX4 expression.","date":"2024","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/38244593","citation_count":19,"is_preprint":false},{"pmid":"25736558","id":"PMC_25736558","title":"Methylene Blue Attenuates iNOS Induction Through Suppression of Transcriptional Factor Binding Amid iNOS mRNA Transcription.","date":"2015","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25736558","citation_count":18,"is_preprint":false},{"pmid":"25244912","id":"PMC_25244912","title":"HUHS1015 induces necroptosis and caspase-independent apoptosis of MKN28 human gastric cancer cells in association with AMID accumulation in the nucleus.","date":"2015","source":"Anti-cancer agents in medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/25244912","citation_count":18,"is_preprint":false},{"pmid":"39971223","id":"PMC_39971223","title":"IGF2BP1/AIFM2 axis regulates ferroptosis and glycolysis to drive hepatocellular carcinoma progression.","date":"2025","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/39971223","citation_count":17,"is_preprint":false},{"pmid":"36845111","id":"PMC_36845111","title":"Lnc-AIFM2-1 promotes HBV immune escape by acting as a ceRNA for miR-330-3p to regulate CD244 expression.","date":"2023","source":"Frontiers in immunology","url":"https://pubmed.ncbi.nlm.nih.gov/36845111","citation_count":17,"is_preprint":false},{"pmid":"32004053","id":"PMC_32004053","title":"Phase Instability amid Dimensional Crossover in Artificial Oxide Crystal.","date":"2020","source":"Physical review letters","url":"https://pubmed.ncbi.nlm.nih.gov/32004053","citation_count":17,"is_preprint":false},{"pmid":"31775552","id":"PMC_31775552","title":"Phages amid antimicrobial resistance.","date":"2019","source":"Critical reviews in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/31775552","citation_count":16,"is_preprint":false},{"pmid":"21559542","id":"PMC_21559542","title":"Hi-Fi transmission of periodic signals amid cell-to-cell variability.","date":"2011","source":"Molecular bioSystems","url":"https://pubmed.ncbi.nlm.nih.gov/21559542","citation_count":16,"is_preprint":false},{"pmid":"28680947","id":"PMC_28680947","title":"Beyond Endemic Burkitt Lymphoma: Navigating Challenges of Differentiating Childhood Lymphoma Diagnoses Amid Limitations in Pathology Resources in Lilongwe, Malawi.","date":"2017","source":"Global pediatric health","url":"https://pubmed.ncbi.nlm.nih.gov/28680947","citation_count":16,"is_preprint":false},{"pmid":"37140233","id":"PMC_37140233","title":"IMPA2 blocks cervical cancer cell apoptosis and induces paclitaxel resistance through p53-mediated AIFM2 regulation.","date":"2023","source":"Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/37140233","citation_count":15,"is_preprint":false},{"pmid":"40187679","id":"PMC_40187679","title":"Deubiquitination by USP7 Stabilizes JunD and Activates AIFM2 (FSP1) to Inhibit Ferroptosis in Melanoma.","date":"2025","source":"The Journal of investigative dermatology","url":"https://pubmed.ncbi.nlm.nih.gov/40187679","citation_count":15,"is_preprint":false},{"pmid":"37839310","id":"PMC_37839310","title":"An updated review on pathogenic coronaviruses (CoVs) amid the emergence of SARS-CoV-2 variants: A look into the repercussions and possible solutions.","date":"2023","source":"Journal of infection and public health","url":"https://pubmed.ncbi.nlm.nih.gov/37839310","citation_count":15,"is_preprint":false},{"pmid":"29857312","id":"PMC_29857312","title":"Neuron-specific regulation of superoxide dismutase amid pathogen-induced gut dysbiosis.","date":"2018","source":"Redox biology","url":"https://pubmed.ncbi.nlm.nih.gov/29857312","citation_count":15,"is_preprint":false},{"pmid":"37735151","id":"PMC_37735151","title":"AIFM2 promotes hepatocellular carcinoma metastasis by enhancing mitochondrial biogenesis through activation of SIRT1/PGC-1α signaling.","date":"2023","source":"Oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/37735151","citation_count":14,"is_preprint":false},{"pmid":"9681529","id":"PMC_9681529","title":"An evaluation of rectal mucosal proliferation measure variability sources in the polyp prevention trial: can we detect informative differences among individuals' proliferation measures amid the noise?","date":"1998","source":"Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/9681529","citation_count":14,"is_preprint":false},{"pmid":"40544552","id":"PMC_40544552","title":"Malnutrition-related diabetes mellitus: Rushing toward \"type 5\" amid unresolved questions and limited evidence.","date":"2025","source":"Diabetes & metabolic syndrome","url":"https://pubmed.ncbi.nlm.nih.gov/40544552","citation_count":13,"is_preprint":false},{"pmid":"34636417","id":"PMC_34636417","title":"Plastid phylogenomics of the Gynoxoid group (Senecioneae, Asteraceae) highlights the importance of motif-based sequence alignment amid low genetic distances.","date":"2021","source":"American journal of botany","url":"https://pubmed.ncbi.nlm.nih.gov/34636417","citation_count":12,"is_preprint":false},{"pmid":"23746973","id":"PMC_23746973","title":"Cell therapy companies make strong progress from October 2012 to March 2013 amid mixed stock market sentiment.","date":"2013","source":"Cell stem cell","url":"https://pubmed.ncbi.nlm.nih.gov/23746973","citation_count":12,"is_preprint":false},{"pmid":"40149908","id":"PMC_40149908","title":"Cardiovascular-Endocrine-Metabolic Medicine: Proposing a New Clinical Sub-Specialty Amid the Cardiometabolic Pandemic.","date":"2025","source":"Biomolecules","url":"https://pubmed.ncbi.nlm.nih.gov/40149908","citation_count":10,"is_preprint":false},{"pmid":"34025772","id":"PMC_34025772","title":"Adaptive markers distinguish North and South Pacific Albacore amid low population differentiation.","date":"2021","source":"Evolutionary applications","url":"https://pubmed.ncbi.nlm.nih.gov/34025772","citation_count":10,"is_preprint":false},{"pmid":"27363647","id":"PMC_27363647","title":"N-acetylcysteine amid reduces pancreatic damage in a rat model of acute necrotizing pancreatitis.","date":"2016","source":"The Journal of surgical research","url":"https://pubmed.ncbi.nlm.nih.gov/27363647","citation_count":10,"is_preprint":false},{"pmid":"28824885","id":"PMC_28824885","title":"AmiD Is a Novel Peptidoglycan Amidase in Wolbachia Endosymbionts of Drosophila melanogaster.","date":"2017","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/28824885","citation_count":9,"is_preprint":false},{"pmid":"39087984","id":"PMC_39087984","title":"Modulation of α-synuclein aggregation amid diverse environmental perturbation.","date":"2024","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/39087984","citation_count":9,"is_preprint":false},{"pmid":"35864848","id":"PMC_35864848","title":"Inhibition of Hippocampal Neurogenesis Starting in Adolescence Increases Anxiodepressive Behaviors Amid Stress.","date":"2022","source":"Frontiers in behavioral neuroscience","url":"https://pubmed.ncbi.nlm.nih.gov/35864848","citation_count":9,"is_preprint":false},{"pmid":"34704616","id":"PMC_34704616","title":"Concerted evolution of ribosomal DNA: Somatic peace amid germinal strife: Intranuclear and cellular selection maintain the quality of rRNA.","date":"2021","source":"BioEssays : news and reviews in molecular, cellular and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34704616","citation_count":9,"is_preprint":false},{"pmid":"30474186","id":"PMC_30474186","title":"Nucleus-localized adiponectin is survival gatekeeper through miR-214-mediated AIFM2 regulation.","date":"2019","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/30474186","citation_count":8,"is_preprint":false},{"pmid":"23230445","id":"PMC_23230445","title":"UniProtKB amid the turmoil of plant proteomics research.","date":"2012","source":"Frontiers in plant science","url":"https://pubmed.ncbi.nlm.nih.gov/23230445","citation_count":8,"is_preprint":false},{"pmid":"38999413","id":"PMC_38999413","title":"Brown Tumors: The Hidden Face of Primary and Renal Hyperparathyroidism Amid Real-Life Settings.","date":"2024","source":"Journal of clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/38999413","citation_count":8,"is_preprint":false},{"pmid":"38776226","id":"PMC_38776226","title":"An evolutionary optimum amid moderate heritability in prokaryotic cell size.","date":"2024","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/38776226","citation_count":7,"is_preprint":false},{"pmid":"37160207","id":"PMC_37160207","title":"Fingolimod mitigates memory loss in a mouse model of Gulf War Illness amid decreasing the activation of microglia, protein kinase R, and NFκB.","date":"2023","source":"Neurotoxicology","url":"https://pubmed.ncbi.nlm.nih.gov/37160207","citation_count":7,"is_preprint":false},{"pmid":"40395523","id":"PMC_40395523","title":"Chaperone-mediated autophagy in fish: A key function amid a changing environment.","date":"2024","source":"Autophagy reports","url":"https://pubmed.ncbi.nlm.nih.gov/40395523","citation_count":7,"is_preprint":false},{"pmid":"37965810","id":"PMC_37965810","title":"Salivary miR-150-5p as an indicator of periodontitis severity and regulator of human periodontal ligament fibroblast behavior by targeting AIFM2.","date":"2023","source":"Journal of periodontal research","url":"https://pubmed.ncbi.nlm.nih.gov/37965810","citation_count":6,"is_preprint":false},{"pmid":"34620344","id":"PMC_34620344","title":"Tailoring nutrition therapy amid the COVID-19 pandemic: Does it work?","date":"2021","source":"Clinical nutrition ESPEN","url":"https://pubmed.ncbi.nlm.nih.gov/34620344","citation_count":6,"is_preprint":false},{"pmid":"31773996","id":"PMC_31773996","title":"Navigating to the most promising directions amid complex fields of vaccine development: a chlamydial case study.","date":"2019","source":"Expert review of vaccines","url":"https://pubmed.ncbi.nlm.nih.gov/31773996","citation_count":6,"is_preprint":false},{"pmid":"39309743","id":"PMC_39309743","title":"Molecular characterization of gliomas and glioneuronal tumors amid Noonan syndrome: cancer predisposition examined.","date":"2024","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/39309743","citation_count":5,"is_preprint":false},{"pmid":"38197738","id":"PMC_38197738","title":"Recognizing facial expressions of emotion amid noise: A dynamic advantage.","date":"2024","source":"Journal of vision","url":"https://pubmed.ncbi.nlm.nih.gov/38197738","citation_count":5,"is_preprint":false},{"pmid":"35997588","id":"PMC_35997588","title":"The Gallus gallus RJF reference genome reveals an MHCY haplotype organized in gene blocks that contain 107 loci including 45 specialized, polymorphic MHC class I loci, 41 C-type lectin-like loci, and other loci amid hundreds of transposable elements.","date":"2022","source":"G3 (Bethesda, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/35997588","citation_count":5,"is_preprint":false},{"pmid":"26637870","id":"PMC_26637870","title":"Phosphatidylinositol Derivatives Induce Gastric Cancer Cell Apoptosis by Accumulating AIF and AMID in the Nucleus.","date":"2015","source":"Anticancer research","url":"https://pubmed.ncbi.nlm.nih.gov/26637870","citation_count":5,"is_preprint":false},{"pmid":"31290091","id":"PMC_31290091","title":"The Impact of Morphine on Reproductive Activity in Male Rats Is Regulated by Rf-Amid-Related Peptide-3 and Substance P Adjusting Hypothalamic Kisspeptin Expression.","date":"2019","source":"Journal of molecular neuroscience : MN","url":"https://pubmed.ncbi.nlm.nih.gov/31290091","citation_count":5,"is_preprint":false},{"pmid":"34257318","id":"PMC_34257318","title":"Malian adults maintain serologic responses to virulent PfEMP1s amid seasonal patterns of fluctuation.","date":"2021","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/34257318","citation_count":5,"is_preprint":false},{"pmid":"33509897","id":"PMC_33509897","title":"Nasopharyngeal diffuse large B-cells lymphoma causing acute airway obstruction amid COVID-19 crisis: an anaesthetist's nightmare.","date":"2021","source":"BMJ case reports","url":"https://pubmed.ncbi.nlm.nih.gov/33509897","citation_count":5,"is_preprint":false},{"pmid":"40990918","id":"PMC_40990918","title":"Human CD4+ T cells recognize Mycobacterium tuberculosis-infected macrophages amid broader responses.","date":"2025","source":"The Journal of experimental medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40990918","citation_count":4,"is_preprint":false},{"pmid":"39365735","id":"PMC_39365735","title":"Dynamic transition of Tregs to cytotoxic phenotype amid systemic inflammation in Graves' ophthalmopathy.","date":"2024","source":"JCI insight","url":"https://pubmed.ncbi.nlm.nih.gov/39365735","citation_count":4,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":46576,"output_tokens":4991,"usd":0.107297,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13365,"output_tokens":5097,"usd":0.097125,"stage2_stop_reason":"end_turn"},"total_usd":0.204422,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2002,\n      \"finding\": \"AMID (AIFM2) is a mitochondrion-associated flavoprotein that induces caspase-independent apoptosis independent of p53 and not inhibited by Bcl-2; it lacks a mitochondrial localization sequence but co-localizes with mitochondria as shown by immunofluorescent staining and biochemical fractionation.\",\n      \"method\": \"Immunofluorescent staining, subcellular fractionation, overexpression in cells, caspase activity assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal localization and functional data from overexpression with multiple readouts, single lab\",\n      \"pmids\": [\"11980907\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"AMID (AIFM2) is a flavoprotein oxidoreductase that binds the unusual modified cofactor 6-hydroxy-FAD (stoichiometrically), possesses NAD(P)H oxidase activity, and catalyzes NAD(P)H-dependent reduction of cytochrome c and molecular oxygen; NADPH binds ~10-fold more tightly than NADH. AMID also binds DNA without apparent sequence specificity, and coenzyme and DNA binding are mutually exclusive, with DNA binding inducing a major conformational change.\",\n      \"method\": \"Recombinant protein purification, in vitro enzymatic assays, spectrophotometric cofactor analysis, binding kinetics (Kd determination), DNA-binding assays\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstituted biochemistry with purified recombinant protein, multiple orthogonal in vitro assays, mutagenesis-level cofactor reconstitution, single rigorous study\",\n      \"pmids\": [\"15958387\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"The yeast AMID homologue Ndi1p (internal NADH dehydrogenase) induces apoptosis-like cell death when overexpressed, an effect suppressed by increased respiration; Ndi1p overexpression increases mitochondrial ROS production, and disruption of NDI1 decreases ROS and extends chronological lifespan.\",\n      \"method\": \"Yeast genetics, overexpression/disruption, ROS measurement, chronological lifespan assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function and overexpression with multiple cellular phenotype readouts, single lab, yeast ortholog\",\n      \"pmids\": [\"16436509\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"DNA binding to AMID (AIF-M2) suppresses its redox activity by preventing NAD(P)H coenzyme binding; binding of DNA and coenzyme is mutually exclusive. DNA binding does not perturb the reduction potential of the 6-hydroxy-FAD cofactor but significantly slows reduction kinetics by NAD(P)H. The DNA-binding domain resides in the C-terminal deletion mutant AIF-M2-(Δ319-613), distinct from AIF whose DNA binds the C-terminal D3 domain.\",\n      \"method\": \"Solution binding assays, stopped-flow kinetics, redox titrations, deletion mutagenesis, quantitative ligand binding analysis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — multiple orthogonal biochemical methods with mutagenesis, in vitro reconstitution, rigorous quantitative analysis in single study\",\n      \"pmids\": [\"17711848\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"AMID protein localizes to the cytoplasm, predominantly at the cytoplasmic side of membranes of various organelles with highest concentration at the Golgi; AMID does not translocate to the nucleus during apoptosis (unlike AIF and endonuclease G), and bioinformatic analysis does not detect nuclear localization signals in AMID.\",\n      \"method\": \"Live-cell fluorescence imaging, single-cell colocalization analysis, time-lapse imaging during staurosporine-induced apoptosis, bioinformatic prediction, nuclear translocation analysis\",\n      \"journal\": \"Apoptosis : an international journal on programmed cell death\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — direct live-cell imaging with quantitative colocalization, multiple methods, but finding of no nuclear translocation is a negative result\",\n      \"pmids\": [\"17347867\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The human AMID gene promoter is a direct transcriptional target of p53: p53 binds the AMID promoter in chromatin immunoprecipitation assays, and deletion mutagenesis identified two p53-responsive elements in the promoter.\",\n      \"method\": \"Luciferase reporter assays, chromatin immunoprecipitation (ChIP), deletion mutagenesis, expression arrays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus reporter assay with mutagenesis, single lab\",\n      \"pmids\": [\"15273740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"AMID-deficient mice generated by gene targeting are viable and fertile with no spontaneous tumors and similar fibrosarcoma incidence after MCA inoculation compared to wild-type; AMID-/- embryonic fibroblasts show slightly increased resistance to genotoxin-induced growth arrest. These data indicate AMID is not required for normal development or p53-mediated tumor suppression.\",\n      \"method\": \"Gene targeting (knockout mouse), tumor incidence assays, MEF proliferation and genotoxin-resistance assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean genetic KO in vivo with multiple phenotypic readouts; negative finding for tumor suppression robustly established\",\n      \"pmids\": [\"16186796\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"AMID protein is found associated with the plasma membrane fraction of apoptotic cells (identified by MALDI-TOF mass spectrometry from Pisum sativum lectin-affinity chromatography), suggesting possible glycosylation and translocation to the plasma membrane during apoptosis; in Jurkat T-cells, AMID expression increases after apoptosis induction.\",\n      \"method\": \"Lectin-affinity chromatography, MALDI-TOF mass spectrometry, immunocytochemistry, Western blotting\",\n      \"journal\": \"Apoptosis : an international journal on programmed cell death\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single approach for membrane localization, no direct functional validation of glycosylation\",\n      \"pmids\": [\"18368494\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"AMID (AIFM2) functions as a rotenone-sensitive NADH:ubiquinone oxidoreductase (NDH-2): when reconstituted into bacterial or mitochondrial membranes, N-terminally tagged AMID displays substantial NADH:O2 activity, supports NADH-linked proton pumping, and rescues growth of E. coli lacking both complex I and NDH-2. C-terminally tagged AMID and NADH-binding site mutants fail to show these activities.\",\n      \"method\": \"Membrane reconstitution, NADH:O2 activity assays, proton pumping assays, E. coli complementation (growth rescue), site-directed mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro reconstitution, mutagenesis, and heterologous complementation with multiple orthogonal assays in one study\",\n      \"pmids\": [\"26063804\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Intracellularly transported adenosine induces caspase-independent apoptosis in MCF-7 breast cancer cells by promoting translocation of AMID from the cytosol to the nucleus; AMID knockdown abrogates adenosine-induced cell death.\",\n      \"method\": \"siRNA knockdown, immunofluorescence (AMID localization), caspase activity assays, cell viability assays, adenosine transporter inhibitor studies\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — knockdown with rescue-relevant pathway placement and localization data, single lab\",\n      \"pmids\": [\"22388174\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"A3 adenosine receptor activation mediates apoptosis in RCC4-VHL human renal cancer cells by upregulating AMID mRNA and protein expression and promoting AMID translocation from cytosol to nucleus; knockdown of either A3 receptor or AMID inhibits adenosine-induced apoptosis.\",\n      \"method\": \"siRNA knockdown, RT-PCR, Western blotting, immunofluorescence, MTT assay, TUNEL staining\",\n      \"journal\": \"The Journal of urology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal knockdown epistasis establishes pathway position, single lab\",\n      \"pmids\": [\"23174235\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"AIFM2/FSP1 blocks ferroptosis through ESCRT-III recruitment to the plasma membrane, activating a membrane repair mechanism involving membrane budding and fission; this mechanism is independent of ubiquinol/coenzyme Q10. Genetic inhibition of the AIFM2-ESCRT-III pathway increases anticancer activity of sorafenib in a xenograft mouse model.\",\n      \"method\": \"Genetic inhibition, xenograft mouse model, ferroptosis induction assays (erastin, sorafenib, RSL3), ESCRT-III pathway analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — in vitro and in vivo genetic experiments with mechanistic pathway placement, single lab\",\n      \"pmids\": [\"31964528\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Aifm2 is a lipid droplet-associated NADH oxidoreductase enriched in brown adipose tissue (BAT) that, upon cold or β-adrenergic stimulation, associates with the outer side of the mitochondrial inner membrane. Acting as a mammalian external NADH dehydrogenase (NDE)-like enzyme, Aifm2 oxidizes NADH to maintain high cytosolic NAD+ levels supporting robust glycolysis and transfers electrons to the electron transport chain (ETC) to fuel thermogenesis. Knockout of Aifm2 impairs cold- and diet-induced thermogenesis, oxygen consumption, and uncoupled respiration.\",\n      \"method\": \"Subcellular fractionation, lipid droplet isolation, cold/β-adrenergic stimulation experiments, NAD+/NADH measurements, oxygen consumption assays, KO mouse model, diet-induced obesity model\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — multiple orthogonal biochemical and genetic methods in vivo and in vitro, KO mouse with defined metabolic phenotypes, subcellular localization with functional consequence\",\n      \"pmids\": [\"31952989\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"FSP1 (AIFM2) inhibition by iFSP1 is species-specific; using cellular, biochemical, and computational methods, a species-specific aromatic architecture was identified as critical for target engagement with the human isoform.\",\n      \"method\": \"Biochemical inhibition assays, computational modeling, cellular ferroptosis assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (biochemical + computational + cellular), single lab\",\n      \"pmids\": [\"37080964\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"ATF6 promotes AIFM2 transcription by binding to both the p53 and AIFM2 promoters; the ATF6/p53/AIFM2 pathway drives acinar cell apoptosis in severe acute pancreatitis. ATF6 knockout suppresses AIFM2 expression and reduces apoptosis, while AIFM2 overexpression reestablishes pathological disorder in ATF6-knockout SAP tissues.\",\n      \"method\": \"ChIP-qPCR, luciferase reporter assay, ATF6 knockout mouse model, adenovirus-mediated shRNA knockdown/overexpression, Western blotting, immunofluorescence\",\n      \"journal\": \"Theranostics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and reporter assays establish direct promoter binding, genetic epistasis in vivo, single lab\",\n      \"pmids\": [\"32724472\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"USP7 deubiquitinates and stabilizes the transcription factor JunD, which in turn promotes transcription of AIFM2 (FSP1), enhancing ferroptosis resistance in melanoma. USP7 inhibition leads to JunD degradation and reduced AIFM2 levels, sensitizing melanoma cells to ferroptosis in vitro and in murine xenograft models.\",\n      \"method\": \"Deubiquitinase identification, Co-IP, Western blotting, gene knockdown/inhibitor experiments, xenograft mouse model, ferroptosis assays\",\n      \"journal\": \"The Journal of investigative dermatology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — epistasis chain established with in vitro and in vivo confirmation, single lab\",\n      \"pmids\": [\"40187679\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"AIFM2 promotes HCC metastasis by activating SIRT1/PGC-1α signaling, increasing mitochondrial biogenesis and oxidative phosphorylation; AIFM2 knockdown impairs HCC metastasis in vitro and in vivo, while overexpression enhances it.\",\n      \"method\": \"shRNA knockdown, overexpression, in vitro migration/invasion assays, in vivo metastasis models, Western blotting for SIRT1/PGC-1α pathway components\",\n      \"journal\": \"Oncogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — gain- and loss-of-function with in vivo validation and pathway identification, single lab\",\n      \"pmids\": [\"37735151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"IGF2BP1 stabilizes AIFM2 mRNA via m6A modification, promoting AIFM2 expression; AIFM2 knockdown suppresses glycolysis and enhances ferroptosis in HCC cells, while overexpression has the opposite effect. IGF2BP1 knockdown effects on glycolysis and ferroptosis are rescued by AIFM2 overexpression.\",\n      \"method\": \"MeRIP assay (m6A detection), mRNA stability assay (Actinomycin D), RIP, siRNA knockdown, overexpression, ferroptosis marker measurement (ROS, iron, MDA/GSH/SOD), glycolysis assays, xenograft/metastasis models\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct RNA modification assay (MeRIP) combined with functional rescue experiments, single lab\",\n      \"pmids\": [\"39971223\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"HUHS1015 (naftopidil analogue) induces caspase-independent apoptosis in MKN28 gastric cancer cells associated with increased nuclear localization of AMID, without changing AMID mRNA or total protein levels, indicating post-translational redistribution.\",\n      \"method\": \"Immunofluorescence (nuclear AMID localization), caspase activity assays, cell cycle analysis, flow cytometry, RT-PCR, Western blotting\",\n      \"journal\": \"Anti-cancer agents in medicinal chemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, correlative localization finding without mechanistic dissection of the translocation mechanism\",\n      \"pmids\": [\"25244912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Adenosine upregulates AMID expression and promotes translocation of AMID from cytosol to nucleus in HuH-7 hepatoma cells; AMID knockdown inhibits adenosine-induced caspase-independent apoptosis, and AMID overexpression enhances it.\",\n      \"method\": \"RT-PCR, immunofluorescence, time-lapse GFP imaging, siRNA knockdown, overexpression, TUNEL staining, flow cytometry\",\n      \"journal\": \"Cellular physiology and biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 / Moderate — reciprocal gain and loss of function with localization data, single lab\",\n      \"pmids\": [\"21325820\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"AIFM2 (AMID/FSP1) is a flavoprotein NADH oxidoreductase that binds 6-hydroxy-FAD and functions as an external NADH dehydrogenase (NDH-2): it oxidizes cytosolic NADH to maintain NAD+ levels supporting glycolysis and feeds electrons to the mitochondrial electron transport chain; its transcription is directly activated by p53 and by ATF6 via p53, and its expression is stabilized post-transcriptionally via IGF2BP1-mediated m6A modification; in ferroptosis, AIFM2/FSP1 suppresses lipid peroxidation-driven cell death through plasma membrane repair via ESCRT-III recruitment and through its reductase activity; DNA binding suppresses its oxidoreductase activity by excluding NAD(P)H; in apoptotic contexts, AMID undergoes cytosol-to-nucleus translocation to drive caspase-independent cell death; and USP7-stabilized JunD directly promotes AIFM2 transcription to confer ferroptosis resistance.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"AIFM2 (AMID/FSP1) is a flavoprotein NAD(P)H oxidoreductase that couples cytosolic redox metabolism to cell-fate decisions [#1, #12]. As a purified enzyme it binds the unusual cofactor 6-hydroxy-FAD stoichiometrically, possesses NAD(P)H oxidase activity, and catalyzes NAD(P)H-dependent reduction of cytochrome c and molecular oxygen, with NADPH binding more tightly than NADH [#1]. When reconstituted into membranes it behaves as a rotenone-sensitive NADH:ubiquinone oxidoreductase (NDH-2), supporting NADH:O2 activity and NADH-linked proton pumping and rescuing E. coli lacking complex I and NDH-2 [#8]. Consistent with this enzymology, Aifm2 acts in vivo as a mammalian external NADH dehydrogenase that oxidizes cytosolic NADH to maintain NAD+ for glycolysis and feeds electrons to the electron transport chain; in brown adipose tissue it is lipid-droplet associated and relocates to the outer face of the mitochondrial inner membrane upon cold or β-adrenergic stimulation, and its loss impairs thermogenesis and uncoupled respiration [#12]. AIFM2 also binds DNA without sequence specificity, and DNA and coenzyme binding are mutually exclusive: DNA binding induces a major conformational change and suppresses redox activity by excluding NAD(P)H [#1, #3]. In ferroptosis, AIFM2/FSP1 acts as a suppressor of lipid-peroxidation-driven death, including through recruitment of ESCRT-III to the plasma membrane to drive membrane repair independently of coenzyme Q10 [#11]. Independently, AMID can drive caspase-independent apoptosis: it is a mitochondrion-associated flavoprotein that triggers death not inhibited by Bcl-2 [#0], and in several cancer contexts adenosine signaling upregulates AMID and promotes its translocation from cytosol to nucleus to execute cell death [#9, #19]. Its expression is controlled transcriptionally — the promoter is a direct p53 target [#5], is activated by ATF6 acting via p53 in acute pancreatitis [#14], and is driven by USP7-stabilized JunD to confer ferroptosis resistance in melanoma [#15] — and post-transcriptionally through IGF2BP1-mediated m6A stabilization of its mRNA, which supports glycolysis and ferroptosis resistance [#17]. Genetic deletion of Aifm2 in mice is compatible with normal development and is dispensable for p53-mediated tumor suppression [#6].\",\n  \"teleology\": [\n    {\n      \"year\": 2002,\n      \"claim\": \"Established AMID as a death-inducing mitochondrion-associated flavoprotein, raising the question of how a protein lacking a mitochondrial targeting sequence executes caspase-independent apoptosis.\",\n      \"evidence\": \"Overexpression, immunofluorescence, subcellular fractionation and caspase assays in cells\",\n      \"pmids\": [\"11980907\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Death effector mechanism not defined\", \"Overexpression-based; endogenous role unaddressed\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Placed AMID under direct transcriptional control of p53, linking its expression to tumor-suppressor signaling.\",\n      \"evidence\": \"ChIP, luciferase reporter and deletion mutagenesis of the human AMID promoter\",\n      \"pmids\": [\"15273740\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether p53-driven AMID contributes to apoptosis in vivo untested\", \"Single cell-context\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Defined AMID as a bona fide oxidoreductase, identifying its 6-hydroxy-FAD cofactor and NAD(P)H oxidase activity, and revealing mutually exclusive DNA and coenzyme binding.\",\n      \"evidence\": \"Recombinant protein, in vitro enzymatic and binding-kinetics assays, DNA-binding assays\",\n      \"pmids\": [\"15958387\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological electron acceptor in cells not established\", \"Functional meaning of DNA binding unresolved\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Genetic knockout showed AMID is dispensable for development and p53-mediated tumor suppression, decoupling its in vitro death activity from an essential physiological requirement.\",\n      \"evidence\": \"Knockout mouse with tumor-incidence and MEF genotoxin-resistance assays\",\n      \"pmids\": [\"16186796\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not exclude context-specific or redundant functions\", \"Metabolic phenotypes not examined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Resolved the mechanism by which DNA binding regulates AMID enzymology — DNA slows NAD(P)H reduction and a C-terminal domain mediates DNA binding — establishing a redox/DNA switch.\",\n      \"evidence\": \"Stopped-flow kinetics, redox titrations and deletion mutagenesis\",\n      \"pmids\": [\"17711848\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vivo relevance of DNA-mediated redox suppression unknown\", \"DNA-binding domain structure not solved\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Localization imaging placed AMID predominantly at cytoplasmic membrane faces (highest at Golgi) and reported no apoptotic nuclear translocation, contesting an AIF-like nuclear death mechanism.\",\n      \"evidence\": \"Live-cell imaging and time-lapse colocalization during staurosporine-induced apoptosis\",\n      \"pmids\": [\"17347867\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Negative nuclear-translocation result conflicts with later cancer studies\", \"Stimulus- and cell-type-specific behavior not reconciled\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"The yeast homologue Ndi1p linked NADH dehydrogenase activity to ROS-dependent death and lifespan, framing the redox-versus-death duality conserved across the family.\",\n      \"evidence\": \"Yeast overexpression/disruption, ROS measurement, chronological lifespan assays\",\n      \"pmids\": [\"16436509\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Ortholog inference, not direct AIFM2 data\", \"Mechanism connecting ROS to death not dissected\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Adenosine signaling was shown to upregulate AMID and drive its cytosol-to-nucleus translocation in caspase-independent apoptosis, defining an upstream death-activation pathway.\",\n      \"evidence\": \"RT-PCR, GFP time-lapse imaging, siRNA and overexpression with TUNEL in hepatoma cells\",\n      \"pmids\": [\"21325820\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Nuclear targeting mechanism unknown (no NLS reported)\", \"Nuclear death effector activity undefined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Extended the adenosine-AMID translocation pathway across breast and renal cancer cells and identified A3 adenosine receptor as an upstream node, generalizing the death mechanism.\",\n      \"evidence\": \"siRNA knockdown, immunofluorescence, viability/TUNEL assays and transporter-inhibitor studies\",\n      \"pmids\": [\"22388174\", \"23174235\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular trigger of translocation unresolved\", \"Reconciliation with negative nuclear-translocation report needed\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Reconstitution defined AMID as an NDH-2-type NADH:ubiquinone oxidoreductase capable of proton pumping and heterologous complementation, anchoring its bioenergetic function.\",\n      \"evidence\": \"Membrane reconstitution, NADH:O2 and proton-pumping assays, E. coli complementation, site-directed mutagenesis\",\n      \"pmids\": [\"26063804\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Topology and physiological membrane association in mammalian cells not fully resolved here\", \"Connection to apoptotic function unclear\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified Aifm2 as a lipid-droplet/mitochondrial external NADH dehydrogenase that sustains cytosolic NAD+ for glycolysis and fuels thermogenesis, defining its core metabolic physiology in vivo.\",\n      \"evidence\": \"Subcellular fractionation, NAD+/NADH and oxygen-consumption assays, KO mouse and diet-induced obesity models\",\n      \"pmids\": [\"31952989\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of stimulus-dependent relocalization mechanistically incomplete\", \"Relationship to ferroptosis pool of AIFM2 not addressed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Revealed a CoQ-independent ferroptosis-suppressing mechanism in which AIFM2/FSP1 recruits ESCRT-III for plasma-membrane repair, broadening its anti-ferroptotic role beyond reductase activity.\",\n      \"evidence\": \"Genetic inhibition, ferroptosis induction assays and xenograft model\",\n      \"pmids\": [\"31964528\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular link between AIFM2 and ESCRT-III recruitment undefined\", \"Relative contribution of repair versus reductase activity unquantified\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Showed ATF6 transactivates AIFM2 via p53 to drive acinar apoptosis, embedding AIFM2 in an ER-stress transcriptional axis in pancreatitis.\",\n      \"evidence\": \"ChIP-qPCR, reporter assays, ATF6 KO mouse and shRNA/overexpression epistasis\",\n      \"pmids\": [\"32724472\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus p53-dependent contribution of ATF6 not fully separated\", \"Single disease context\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Characterized species-specific pharmacology of the FSP1 inhibitor iFSP1, identifying an aromatic architecture required for human target engagement, informing therapeutic targeting.\",\n      \"evidence\": \"Biochemical inhibition, computational modeling and cellular ferroptosis assays\",\n      \"pmids\": [\"37080964\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural binding mode not crystallographically resolved\", \"In vivo efficacy across species not tested\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Linked AIFM2 to a pro-metastatic program via SIRT1/PGC-1α-driven mitochondrial biogenesis and oxidative phosphorylation, connecting its redox role to cancer aggressiveness.\",\n      \"evidence\": \"shRNA/overexpression, migration/invasion and in vivo metastasis assays in HCC\",\n      \"pmids\": [\"37735151\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which AIFM2 activates SIRT1/PGC-1α unclear\", \"Whether reductase activity is required not tested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined transcriptional and post-transcriptional control of AIFM2 in ferroptosis resistance — USP7–JunD transactivation and IGF2BP1 m6A mRNA stabilization — coupling AIFM2 expression to glycolysis and cancer cell survival.\",\n      \"evidence\": \"Co-IP, DUB identification, MeRIP/RIP, mRNA-stability and rescue assays with xenograft models\",\n      \"pmids\": [\"40187679\", \"39971223\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether these regulators act on the same AIFM2 pool as the metabolic/ferroptosis function unclear\", \"Direct binding of JunD to AIFM2 promoter mapped only in melanoma\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how a single flavoprotein partitions between its bioenergetic NADH-dehydrogenase role, its membrane-repair/ferroptosis-suppressing role, and its pro-apoptotic nuclear-translocation behavior, and what governs that partitioning.\",\n      \"evidence\": \"No timeline discovery reconciles the metabolic, ferroptotic, and apoptotic activities mechanistically\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model integrating DNA-binding, membrane-association and reductase states\", \"Conflicting nuclear-translocation reports unreconciled\", \"Trigger for context-specific functional switching unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016491\", \"supporting_discovery_ids\": [1, 8, 12]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [1, 3]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [0, 12]},\n      {\"term_id\": \"GO:0005811\", \"supporting_discovery_ids\": [12]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [11]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [4, 9]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [9, 19]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 11]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [12, 8]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"ESCRT-III\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":9,"faith_total":9,"faith_pct":100.0}}